Value of drug resistance and homology of Acinetobacter baumannii in tracing the source of nosocomial infection in Jianyang, Sichuan / 中国热带医学

@#Abstract: Objective　To analyze the drug sensitivity and the carrying of carbapenem resistant gene of Acinetobacter baumannii isolated from clinical patients and clinical objects, and analyze the homology of strains to provide support for the control of nosocomial infection. Methods　A total of 38 strains of Acinetobacter baumannii isolated from patients and clinical objects surface were collected from January 2019 to August 2020. The antimicrobial susceptibility was tested by the minimum inhibitory concentration method. In addition, the resistance related genes were detected by polymerase chain reaction method, and homology analysis was performed by enterobacterial repetitive Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR). Results All 34 strains of Acinetobacter baumannii isolated from Clinical patients and 4 strains isolated from clinical objects carried blaOXA-51 and imp resistance genes, neither of them carried blaVIM gene. 32 Acinetobacter baumannii carrying blaOXA-23 gene, 28 strains carrying blaTEM gene, 7 strains carrying blaOXA-58 gene. After cluster analysis, 38 Acinetobacter baumannii isolates were classified into 7 genotypes (expressed A, B, C, D, E, F, G), and cluster E and cluster G were the main clusters, containing 12 strains (12/38, 31.6%) and 18 strains (18/38, 47.4%), respectively, as the main prevalent clonal strains. Conclusions　Acinetobacter baumannii isolated from different sources have the significant differences in drug resistance and carry different resistance genes. There is no direct correlation between patients and environmental isolates of Acinetobacter baumannii belonging to different clonal strains. Also, there aren’t significant correlation between clinical patients infected with Acinetobacter baumannii.

Preparation and preliminary evaluation of sodium alginate/agarose/carboxymethyl chitosan spray film dressing carrying platelet rich plasma for wound repair / 中国输血杂志

【Objective】 To develop a spray-on membrane dressing for wound repair containing platelet rich plasma (PRP) sodium alginate (SA)/agarose(AG)/carboxymethyl chitosan (CMCS). 【Methods】 SA/AG/ CMCS were mixed in different proportions to prepare biodegradable quick setting spray (BQSS) by blending film method, and the film-forming time, moisture retention and compression resistance of the prepared BQSS were tested. Then PRP and BQSS were mixed in the proportion of 3∶7, 4∶6, 5∶5, 6∶4 and 7∶3 to prepare PRP-BQSS spray film dressings. The film-forming time, moisture retention, compressive strength, porosity and slow-release effect of growth factors of PRP-BQSS spray film dressings were studied. 【Results】 In the preparation of BQSS compound spray film solution, when SA, AG, CMCS and sterile distilled water were 0.6∶0.6∶0.6∶98.2g, the film-forming time (7.73±0.31) s, moisture retention (75. 54±3.03) % and compression resistance (791.00±68.02) g of the spray-film dressing were the best. The basic properties of PRP-BQSS spray-on film dressings and the release of growth factors show that PRP-BQSS spray-on film dressings can exist in different forms, and with the decrease of PRP concentration percentage, its film-forming time, moisturizing performance and compressive strength showed an upward trend. When the PRP content is 30%, the porosity of the dressing is the highest, about(84.34±0.90)%. The release of platelet-derived growth factor-AA(PDGF-AA), platelet factor-4(PF-4) and transforming growth factor beta (TGF-β) was in a slow upward trend, and the release of the three growth factors was higher than that of PRP group in 48 hours. 【Conclusion】 The preparation method of PRP-BQSS spray film dressing designed in this study is simple and mild, and can form a film quickly, with good biological properties and better growth factor inhibition and sustained-release effect.

Dentin slice model of dental stem cells in a fibrin-agarose construct for dental pulp regeneration / Modelo de trozo de dentina de células madre dentales en una construcción de fibrina-agarosa para la regeneración de la pulpa dental

Objectives: To implement a dentin slice model of mesenchymal stem cells derived from dental tissues in a fibrin-agarose construct for dental pulp regeneration. Material and Methods: MSCs derived from different oral cavity tissues were combined with a fibrin-agarose construct at standard culture conditions. Cell viability and proliferation tests were assayed using a fluorescent cell dye Calcein/Am and WST-1 kit. The proliferation assay was evaluated at 24, 48, 72, and 96 hours. Also, we assessed the dental pulp stem cells (DPSCs) cell morphology inside the construct with histological stains such as Hematoxylin and Eosin, Masson's trichrome, and Periodic acid­Schiff. In addition, we elaborated a tooth dentin slice model using a culture of DPSC in the fibrin­agarose constructs co-adhered to dentin walls. Results: The fibrin-agarose construct was a biocompatible material for MSCs derived from dental tissues. It provided good conditions for MSCs' viability and proliferation. DPSCs proliferated better than the other MSCs, but the data did not show significant differences. The morphology of DPSCs inside the construct was like free cells. The dentin slice model was suitable for DPSCs in the fibrin-agarose construct. Conclusion: Our findings support the dentin slice model for future biological use of fibrin-agarose matrix in combination with DPSCs and their potential use in dental regeneration. The multipotency, high proliferation rates, and easy obtaining of the DPSCs make them an attractive source of MSCs for tissue regeneration.

Objetivos: Implementar un modelo de dentina con células madre mesenquimales derivadas de tejidos dentales en una constructo de fibrina-agarosa para la regeneración de la pulpa dental. Material y Métodos: Las MSC derivadas de diferentes tejidos de la cavidad oral se combinaron con una construcción de fibrina-agarosa en condiciones de cultivo estándar. Las pruebas de viabilidad y proliferación celular se ensayaron utilizando un kit de colorante celular fluorescente Calcein/Am y WST-1. El ensayo de proliferación se evaluó a las 24, 48, 72 y 96 horas. Además, evaluamos la morfología celular de las células madre de la pulpa dental (DPSC) dentro de la construcción con tinciones histológicas como hematoxilina y eosina, tricrómico de Masson y ácido peryódico de Schiff. Además, elaboramos un modelo de rebanadas de dentina dental utilizando un cultivo de DPSC en las construcciones de fibrina-agarosa coadheridas a las paredes de la dentina. Resultados: La construcción de fibrina-agarosa fue un material biocompatible para las MSC derivadas de tejidos dentales. Proporcionó buenas condiciones para la viabilidad y proliferación de las MSC. Las DPSC proliferaron mejor que las otras MSC, pero los datos no mostraron diferencias significativas. La morfología de las DPSC dentro de la construcción era como la de las células libres. El modelo de corte de dentina fue adecuado para DPSC en la construcción de fibrina-agarosa.Conclusión: Nuestros hallazgos respaldan el modelo de corte de dentina para el futuro uso biológico de la matriz de fibrina-agarosa en combinación con DPSC y su uso potencial en la regeneración dental. El multipotencial, las altas tasas de proliferación y la fácil obtención de las DPSC las convierten en una fuente atractiva de MSC para la regeneración de tejidos.

Amino Acid Supplementation Improves the Production of Extracellular Peptidases by Aspergillus Section Flavi and their Ionic Immobilization

Abstract Bioprocess studies have been highlighted due to the importance of physiological processes and industrial applications of enzymes. The potential of peptidase production from Aspergillus section Flavi using different amino acids as a supplemental nitrogen source was investigated. A production profile revealed that amino acids had positive effects on peptidase production when compared to the control without amino acids. Optimal production (100 U/mL) was obtained with Arginine amino acid in 96 h of fermentation. Extracellular peptidase from Aspergillus section Flavi was identified in submerged bioprocesses by in situ activity. Biochemical studies revealed that the maximum activities of the enzyme extract were obtained at pH 6.5 and a temperature of 55°C. The inhibition by EDTA and PMSF suggests the presence of more than one peptidase while the Ni2+ and Cu2+ had a negative influence on the enzyme activity. When the crude extract was reversibly immobilized on ionic supports, DEAE-Agarose and MANAE-Agarose the derivative showed different profiles of thermal and pH stabilities. Hence, this study revealed the basic properties and biochemical characteristics that allowed the production improvement of this class of enzyme. Moreover, with known properties stabilization and immobilization process is required to further explore its biotechnological capacities.

Effect of Stains on the Purity Determination of Human Serum Albumin by Agarose Gel Electrophoresis / 中国药学杂志

OBJECTIVE: To study the effect of stains on the purity detection results of human serum albumin by agarose gel electrophoresis, thus to select appropriate stain so that the detection result of agarose gel electrophoresis method can be basically consistent with the method specified in the 2015 edition of Chinese Pharmacopoeia, cellulose acetate membrane electrophoresis method. METHODS: Acid blue, amino black and ponceau were used in agarose gel electrophoresis to detect the purity of 30 batches of human serum albumin samples, and the results were compared with those obtained by cellulose acetate membrane electrophoresis. RESULTS: Impurity bands could not be detected by ponceau stain; the results of amino black staining was significantly lower than that of cellulose acetate membrane electrophoresis. There was no statistical difference between the results of amino black staining and cellulose acetate membrane electrophoresis. The results of 25 batches of samples detected by cellulose acetate membrane electrophoresis were within the 95% confidence interval of the results detected by amino black staining. CONCLUSION: The category of stain has great influence on the detected purity of human serum albumin when using agarose gel electrophoresis. The results of amino black staining is consistent with those of the method recorded in the Chinese Pharmacopoeia in use.

Preparation of immune microarray carrier based on agarose self-assembled membrane and its application in probe immobilization / 中国免疫学杂志

Objective:To improve the immobilization efficiency of antibody molecules on immune microarray,the method of es-tablishment and optimization of agarose self-assembled membrane carrier with three-dimensional hydrogel structure was established. Methods: The agarose self-assembled membrane carrier was prepared by using glass slide as the carrier,using agarose and sodium periodate modification on glass surface. The agarose self-assembled membrane carrier was characterized by TEM, AFM and FTIR. The optimum preparation conditions were obtained. The carrier for two different species of fixed source antibody efficiency were studied. Antibody loading capacity of agarose self-assembled membrane carrier and ordinary aldehyde carrier were investigated and compared by fluorescence microscopy imaging and Image J software. Results: The agarose nano-membrane carrier had uniform and compact surface. This structure could increase the specific surface area and improve the probe fixed rate. The optimal concentration of agarose for preparation of carrier was 1. 0% . When the concentration of IgG was 0. 3-0. 4 mg/ml,the oxidized self-assembled chitosan film substrate had highest antibody loading capacity. And it had a 3. 94 fold higher antibody loading capacity than the ordinary aldehyde carrier. Conclusion: The agarose nano-membrane carrier is an ideal method for surface modification of immobilized antibody molecules, which is more suitable for preparation of immune microarray carrier.

Evaluation of caprine arthritis-encephalitis virus transmission in newborn goat kids / Estudo da transmissão do vírus da artrite-encefalite caprina em cabritos neonatos

Caprine arthritis encephalitis causes considerable losses in goat production. The main form of the caprine arthritis encephalitis virus transmission is through the ingestion of colostrum or milk from infected females. However, some transmissions cannot be explained in this manner. Therefore, this study aimed to evaluate transplacental transmission of caprine arthritis encephalitis virus. Blood samples were collected from 283 newborn kids of Anglo-Nubian and Saanen breeds born from seropositive and seronegative goats. Samples were collected immediately after birth and analyzed with agarose gel immunodiffusion and western blot. All samples were negative in the agarose gel immunodiffusion. However, the western blot test demonstrated that four kids were born positive for caprine arthritis encephalitis virus. This result indicates that although in a low frequency (1.4%), there is a possibility of transplacental transmission of small ruminant lentivirus.(AU)

A artrite encefalite caprina causa perdas consideráveis para a produção caprina. A principal forma de transmissão do vírus da artrite encefalite caprina é a ingestão de colostro ou leite de fêmeas infectadas. No entanto, algumas transmissões não podem ser explicadas por esta via. Dessa forma, este estudo teve como objetivo avaliar a transmissão do vírus da artrite encefalite caprina por via transplacentária (vertical). Foram realizadas coletas de sangue em 283 crias recém-nascidas das raças Anglo-Nubiana e Saanen, provenientes de progenitores soropositivos e soronegativos. As amostras foram coletadas logo após o nascimento e analisadas pelas técnicas de imunodifusão em gel de agarose e western blot. No teste de imunodifusão em gel de agarose, nenhum cabrito foi detectado reagente. Porém, no teste de western blot, quatro cabritos nasceram soropositivos. Esse resultado indica que, apesar de baixa frequência (1,4%), existe a possibilidade de transmissão via transplacentária do lentivírus de pequenos ruminantes.(AU)

Influence of cell free fetal fraction DNA for noninvasive prenatal test / 国际检验医学杂志

Objective To explore the influence of lower concentration of cell free fetal fraction DNA in maternal plasma on non-invasive prenatal test(NIPT) .Methods A total of 3240 pregnant women accepted NIPT in Foshan Maternal and Children′s Hos-pital from April ,2015 to March ,2016 were analyzed retrospectively ,and 150 samples of which were male fetus judged by Z score of Y chromosome and the cell free fetal fraction DNA were lower than 8% were selected .The cell free fetal fraction DNA were in-creased by agarose gel electrophoresis ,then conducted NIPT ,compared with the results of aneuploidy screening .Results The cell free fetal fraction DNA were increased from 5% to 9 .2% by agarose gel electrophoresis .The result of NIPT after increasing fetal fraction was consistent with it before .Conclusion Concentration of cell free fetal fraction DNA has no influence on the result of NIPT when cell free fetal fraction DNA is above 5% .

Influence of cell free fetal fraction DNA for noninvasive prenatal test / 国际检验医学杂志

Objective To explore the influence of lower concentration of cell free fetal fraction DNA in maternal plasma on non-invasive prenatal test(NIPT) .Methods A total of 3240 pregnant women accepted NIPT in Foshan Maternal and Children′s Hos-pital from April ,2015 to March ,2016 were analyzed retrospectively ,and 150 samples of which were male fetus judged by Z score of Y chromosome and the cell free fetal fraction DNA were lower than 8% were selected .The cell free fetal fraction DNA were in-creased by agarose gel electrophoresis ,then conducted NIPT ,compared with the results of aneuploidy screening .Results The cell free fetal fraction DNA were increased from 5% to 9 .2% by agarose gel electrophoresis .The result of NIPT after increasing fetal fraction was consistent with it before .Conclusion Concentration of cell free fetal fraction DNA has no influence on the result of NIPT when cell free fetal fraction DNA is above 5% .

Application of modified agarose electrophoresis for detecting bone alkaline phosphatase isoenzyme in diagnosis of bone metastasisof prostate carcinoma / 国际检验医学杂志

Objective To discuss the application of improved agarose electrophoresis for detecting bone alkaline phosphatase(BALP) isoenzymes in the diagnosis of metastasis in prostate carcinoma(PCa).Methods Serum ALP isoenzymes in 50 persons undergoing physical examination and 163 cases of PCa were detected by the improved agarose electrophoresis after treating serum by neuraminidase.Results (1)The resolution of liver and bone ALP by the common agarose gel electrophoresis was lower,and which was increased by the modified agarose electrophoresis.(2)Total ALP and BALP in the PCa group were higher than those in the control group;total ALP and BALP activity in the PCa bone metastasis group were higher than those in the control group and non-metastasis group(P0.05).BALP and total ALP in bone metastasis of prostate carcinoma were higher than no bone metastasis.Conclusion Bone ALP has a certain correlation with PCa bone metastasis,also between bone ALP and bone metastasis of carcinoma.After treating serum by neuraminidase,adopting the improved agarose gel electrophoresis for separating ALP isozymes contributes to the clinical auxiliary diagnosis of PCa bone metastasis.

Principios y relevancia del ensayo cometa / Principles and relevance of the comet assay

Desde hace varias décadas el ensayo Cometa, o electroforesis alcalina de células individuales, se ha convertido en un método establecido para el estudio del daño de ácido desoxirribonucleico, con múltiples aplicaciones en ensayos de genotoxicidad, estudios de biomonitoreo en humanos, epidemiologia molecular y ecotoxicología; así como una herramienta fundamental para investigaciones sobre daño y reparación del ácido desoxirribonucleico. Este ensayo se distinguió por su simplicidad, sensibilidad, versatilidad, rapidez y economía. Es una poderosa técnica que se basa en la visualización microscópica de las imágenes del ácido desoxirribonucleico después que las células son embebidas en agarosa, lisadas y sometidas a una electroforesis alcalina. Esta metodología básica ha sido ampliada, y permite ahora, detectar con alta sensibilidad una gran variedad de daños del ácido desoxirribonucleico en cualquier tipo de células. La inclusión en este ensayo, de enzimas capaces de producir lesiones específicas en la hebra de ácido desoxirribonucleico, ha incrementado su rango de detección y sensibilidad. Pero es importante tener claro que su especificidad no es absoluta. El propósito es destacar algunos aspectos útiles de este método y sus ventajas; describir la experiencia en algunos aspectos técnicos del proceder, normalizado según las condiciones del laboratorio en el instituto para ampliar su utilización en el país.

For several decades now the comet assay (single cell gel electrophoresis assay) has been the method used for the study of deoxyribonucleic acid damage, with multiple applications in genotoxicity assays, biomonitoring studies in humans, molecular epidemiology and ecotoxicology, and a fundamental tool for research into deoxyribonucleic acid damage and repair. The comet assay has stood out for its simplicity, sensitivity, versatility, rapidity and economy. It is a powerful technique based on microscopic visualization of deoxyribonucleic acid images after the cells have been embedded in agarose, lysed and subjected to alkaline electrophoresis. This basic methodology has been broadened, and may now detect with great sensitivity a large variety of deoxyribonucleic acid damage in any type of cell. Inclusion in the assay of enzymes capable of producing specific lesions on the deoxyribonucleic strand has broadened its detection range and sensitivity. However, it is important to bear in mind that its specificity is not absolute. The purpose of the present study is to point out some useful aspects and advantages of the method, and describe the experience with some technical aspects of the procedure, standardized in keeping with the conditions in the laboratory at the institute to extend its use in the country.

Variantes de hemoglobina en una población con impresión diagnóstica positiva para hemoglobinopatías en Colombia / Hemoglobin variants in Colombian patients referred to discard hemoglobinopathies

Background: Oxygen transport is altered in hemoglobinopathies. Aim: To study the distribution of hemoglobinopathies in Andean subjects without African ancestry. Material and Methods: We analyzed blood samples of 1,407 subjects aged 18 to 59 years (58% females), living in the central Andean region of Colombia, referred to discard hemoglobinopathies. The frequency and type of hemoglobinopathy was established by capillary and agarose gel electrophoresis. Results: The frequency of hemoglobinopathies was 34.5% and higher among females. The structural variants found were: AS-heterozygous hemoglobin (8.1%), homozygous SS (3.7%), heterozygous SC (2.2%), AC heterozygotes (0.5%) and heterozygous AE (0.3%). Quantitative variants found were Hb A-Beta thalassemia (13.91%) and Hb H (0.06%), Beta-thalassemia heterozygotes C (0.88%), S-Beta thalassemia heterozygotes (6.07%) and compound heterozygous SC/Beta thalassemia (0.25%), with a persistence of fetal hemoglobin 0. Composite thalassemia was also found in 31%. All techniques showed good correlation and capillary electrophoresis demonstrated a greater detection of hemoglobin variants. Conclusions: The frequency of hemoglobin variants in the analyzed population was high, which is an important public health indicator. The most common hemoglobin variant was HbA/Increased structural Hb A2 and the mos frequent structural hemoglobinopathy was sickle cell trait. Capillary electrophoresis can discern any Hb variants present in the population.

Searching for the best agarose candidate from genus Gracilaria, Eucheuma, Gelidium and local brands

Objective: To explore the potential of local agar of genus Gracilaria, Eucheuma, Gelidium and local brands as an alternative for imported agarose for DNA electrophoresis, and to examine their ability related to separation and migration of DNA fragments in DNA electrophoresis. Methods: Their performance at various concentrations were compared via an experimental study with a specific brand of imported commercial agarose used in molecular biology research. The measured variables were separation and migration during electrophoresis of a DNA fragment. Results: The local agar genus Gracilaria gigas, Gelidium, brand "B" and brand "S" could separate DNA fragments at a concentration between 1% and 2%, with an optimum concentration of 2% w/v, as good as a specific brand of imported commercial agarose. Conclusions: Their performance were very close to that of commercial agarose and can still be improved by further agar purification as well as by pH and sulfur control.

Electrophoretic Techniques for the Detection of Human Microsatellite D19S884

Background: The detection and analysis of microsatellites is very important for the mapping of genetic diseases because they are commonly used as genetic markers. Microsatellite marker D19S884 has been associated with polycystic ovary syndrome (PCOS), the most common reproductive endocrine disease of women in their childbearing years. It is responsible for an estimated 70% of cases of anovulatory infertility. In this work, we detected microsatellites in DNA extracted from the blood of PCOS patients. Methods: DNA microsatellites were amplified by polymerase chain reaction (PCR) using a pair of specific primers tagged with fluorescence to yield products of 160–200 base pairs in length. Alleles were separated on 4% low-melting agarose gels; stained with a safe gel staining, GelRed™, which is an alternative to ethidium bromide; and visualised by ultraviolet illumination. Results: Bands were observed, but their base-pairs differences were difficult to distinguish. To identify each allele clearly, the PCR products were also analysed using capillary gel electrophoresis for fragment analysis where it was possible to discriminate even in case of difference between two pairs of bases between the alleles. Conclusion: In this article, we present a protocol that combines the use of gel electrophoresis and fragment analysis in the identification of genetic biomarkers for PCOS.

Searching for the best agarose candidate from genusGracilaria,Eucheuma,Gelidium and local brands

Objective:To explore the potential of local agar of genusGracilaria,Eucheuma,Gelidium and local brandsas an alternative for imported agarose forDNA electrophoresis, and to examine their ability related to separation and migration ofDNA fragments inDNA electrophoresis. Methods:Their performance at various concentrations were compared via an experimental study with a specific brand of imported commercial agarose used in molecular biology research. The measured variables were separation and migration during electrophoresis of a DNA fragment. Results: The local agar genusGracilaria gigas,Gelidium, brand B and brand S could separateDNA fragments at a concentration between 1% and 2%, with an optimum concentration of 2% w/v, as good as a specific brand of imported commercial agarose. Conclusions:Their performance were very close to that of commercial agarose and can still be improved by further agar purification as well as by pH and sulfur control.

Analysis and significance of results of hemoglobin electrophoresis in 7 652 cases / 国际检验医学杂志

Objective To investigate the detection rates and types of hemoglobinopathy in Xiaolan district of Zhongshan city to provide the basis for prepotency .Methods 7 652 pregnant women with pregnancy detection in our hospital from June 2010 to De‐cember 2013 were selected and performed the electrophoresis by the Hydrasys automatic hemoglobin electrophoresis system .The hemoglobin components were scanned out and performed the statistical analysis .Results Among 7 652 samples ,1 057 cases of sus‐pected hemoglobinopathy were screened out with the detection rate of 10 .36% .Among them ,511 cases(6 .68% ) were Hb A2 3 .5% (suspected β‐thalassemia) ,the detection rate of Hb A2 >3 .5% was lower than that of Hb A2 < 2 .0% ,the difference had statistical significance(P< 0 .05) .11 cases (0 .14% ) were Hb C or E ,9 cases (0 .12% ) were Hb J or K ,8 cases (0 .10% ) were Hb H ,7 cases (0 .09% ) were Hb Bart′s ,1 case was Hb CS ,2 cases were Hb G and 1 case was Hb D ,which accounted for 0 .04% .Conclusion Hemoglobin electrophoresis can screen hemoglobinopa‐thy effectively .The hemoglobinopathy screening in this area should be conducted as soon as possible ,which has an important signif‐icance to prepotency .

Synthesis of nonviral gene carrier poly (L-glutamic acid)-graft-polypropylenimine and its complexation stability with DNA / 中国药学杂志

OBJECTIVE: To synthesize nonviral gene carrier poly(L-glutamic acid)-graft-polypropylenimine(MP-g-PPI) and investigate the capability of the polymers to condense DNA and the stability of MP-g-PPI DNA complexes.

Polyclonal all immune globulin hematic disease laboratory appraisal analysis discussed in this paper / 国际检验医学杂志

Objective To explore the clinical diagnostic value of polyclonal all immunoglobulin.Methods The specimens of pa-tients were simultaneously tested and identified by quantitative immunoglobulins,Immunofixation electrophoresis of serum and u-rine,urine protein electrophoresis,and other ways.Results From 1 patients with rheumatoid arthritis were detected the serum IgG, M,A,KAP and LAM,and urine KAP and LAM,at the same time show the increment of the polyclonal polyclonal all immune glob-ulin hematic disease.Conclusion Polyclonal all immune globulin hematic disease often appear in the complications of chronic in-flammation,which should be paid attention during its in clinical doctors.

Application of improved gel electrophoresis to the detection of genotype polymorphisms / 中华实用儿科临床杂志

Objective To explore the traits of improved polyacrylamide gel electrophoresis (PAGE)-ethidium bromide(EB) staining in the detection of genotype polymorphisms.Methods The methods of PAGE-silver staining,agarose gel electrophoresis (AGE)-EB staining and improved PAGE-EB staining in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the μ-opioid receptor (OPRM1) A118G genotypes in case group (n =167) infants with intracranial hemorrhage (ICH),and control group(n =163) infants without ICH,to conduct a case study analysis.And the application traits of three methods were compared.Results Genotypes of OPRM1 A118G were GG (169 bp),AG (193 bp,169 bp),AA (193 bp).Both the electrophoresis methods of PAGE-silver staining and PAGE-EB could be used to detect the genotypes of OPRM1 A118G clearly in this study.There was no statistically significant difference between the resolutions of DNA fragments (P =0.884).The first method,which had 13 experiment steps,consuming 4-5 hours,involving 12 kinds of chemical reagents,and the pictures were taken with the camera,was complex,with difficult operation,more time consuming; Compared with the first method,the secondmethod was simple,which had 6 test procedures,consuming 2 hours with 8 reagents,and the pictures were taken by using an automatic gel imager.AGE-EB could not be used to detect genotypes of OPRM1 A118G.Conclusions The method of improved PAGE-EB has the advantages of fast,easy operation,and high resolution,which is worthy of wider application.

Curing effects evaluation of rhizoma atractylodis macrocephalae on yeast prion[PSI+] / 中国人兽共患病学报

Investigation of the effects of rhizome atractylodis macrocephalae on yeast prion [PSI+ ]was conducted in this study .Liquid culture containing rhizome atractylodis macrocephalae aqueous extracts was applied to evaluate its therapeutic effects preliminarily .Then yeast replica plating together with Semi Denaturing Detergent Agarose Gel Electrophoresis com-bined with western blot were used to further confirm the results .It's showed that the cure rate of aqueous extracts of rhizome atractylodis macrocephalae on yease prion [PSI+ ]could reach 6% .